skoufos at nefeli.imbb.forth.gr wrote:
: Dear Netters:
: Does any of you know of expression vectors that release the expressed
: protein in the tissue culture medium and work in mammalian cell lines?
: Or do you know of suitable candidates to be used as fussion proteins
: for construction of such vectors?
I have just started using secreted alkaline phosphatase (SEAP) in
transfections of a variety of cell types and it has worked pretty well,
with a sensitivity nearly as good as luciferase using a luminiscent
substrate. I got the vector and detection buffers from Tropix, I think
that Clontech also sells it (I think they call it "The Great Escape SEAP
kit", or something equally silly). So far I like it because I only need
to use about 0.5 ug/transfection (my cells don't like to overdose on DNA,
which they were with my B-Gal vector) and get good results. Hope this is
helpful,
Sheryl
----------------------- Sheryl L White -------------------------
Dept of Mol Physiology, Given Med, UVM, Burlington, VT 05405
Voice: (802) 656-4433 E-Mail: slw at salus.med.uvm.edu
----------------------------------------------------------------